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Identification of pure and mixed basophil colonies in culture of human
peripheral blood and marrow cells
AG Leary and M Ogawa
We present a colony assay system that allows in situ identification of
human basophil/mast cell (basophil) colonies. In methylcellulose culture,
in the presence of phytohemagglutinin-leukocyte conditioned media
(PHA-LCM), human peripheral blood and bone marrow cells form colonies that
can be distinguished by their unique morphological characteristics. Pure
basophil colonies are diffuse, small colonies containing small, round,
highly refractile cells. These characteristics of the constituent cells led
us to the observation that a significant number of basophils are found in
combination with eosinophils. The mixed eosinophil/basophil colonies have
the distinctive elements of pure eosinophil and pure basophil colonies.
Usually, these are diffuse colonies with compact clusters of slightly
larger, darker-appearing cells. We also found colonies that contained
basophils and neutrophils/monocytes, but this type could not be
consistently identified by in situ morphology. Cytochemical analysis
confirmed the metachromatic nature of the granules in the basophils. The
presence of IgE receptors on the cells was documented by indirect
immunofluorescent staining after passive sensitization with purified human
IgE. Peripheral blood cells from six healthy volunteers formed 5.7 +/- 1.0
(mean +/- SEM) pure colonies in 2 X 10(5) cells. Cultures of bone marrow
cells from patients with various types of anemia had 9.0 +/- 1.5 colonies
in 10(5) cells. This is the first description of a colony assay system for
in situ identification of a pure population of basophilic granulocytes.
Volume 64,
Issue 1,
pp. 78-83,
07/01/1984
Copyright © 1984 by The American Society of Hematology
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